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Application of stress resistance-related protein bhlh85 in regulating plant stress resistance

A plant and salt-resistant technology, applied in the application field of stress-resistance-related protein bHLH85 in regulating plant stress-resistance, which can solve the problems of little research on bHLH transcription factors.

Active Publication Date: 2022-05-13
SHANDONG NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the inventors found that at present, the research is mainly on a small number of model organisms such as Arabidopsis, and there is little research on bHLH transcription factors in other plants

Method used

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  • Application of stress resistance-related protein bhlh85 in regulating plant stress resistance
  • Application of stress resistance-related protein bhlh85 in regulating plant stress resistance
  • Application of stress resistance-related protein bhlh85 in regulating plant stress resistance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Example 1: Bioinformatics analysis of sweet sorghum bHLH85 gene

[0065] 1 Experimental materials

[0066] Sweet sorghum (Sb08g019780), and Japanese rice (LOC107276154), castor (LOC8272269), upland cotton (LOC107930558), corn (LOC103653155), squash (LOC111455548), morning glory (LOC109169792), cassava ( LOC110618953), the HLH domain sequence of Arabidopsis (AT4G33880) and other proteins.

[0067] The data comes from the bHLH85 protein of the sweet sorghum salt-tolerant high-sugar inbred line M-81E on the NCBI website and the BLAST results of the bHLH85 domain of other species.

[0068] 2 Experimental methods

[0069] First, find the cDNA sequence of sweet sorghum gene bHLH85 on the NCBI website, translate it into amino acid sequence, and then use ExPASy software to analyze the primary structure and hydrophobicity of the protein according to the amino acid sequence; Level structure analysis; use TMHMM software to predict and analyze the transmembrane domain of the pro...

Embodiment 2

[0088] Example 2 Cloning, Vector Construction and Subcellular Localization of Sweet Sorghum bHLH85 Gene

[0089] 1 Experimental materials and material handling

[0090] 1.1 Experimental materials

[0091] Sweet sorghum salt-tolerant high-sugar inbred line M-81E seeds (new seeds from the previous year), Nicotiana benthamiana seeds, pEASY-Blunt3 simple cloning vector, pROKII-GFP expression vector (driven by CaMV35S promoter), DH5α Escherichia coli strains, GV3101 Agrobacterium strain, etc.

[0092] 1.2 Material Handling

[0093] The seeds of sweet sorghum salt-tolerant inbred line M-81E with the same size, complete and plump seeds were selected, placed in mesh bags, and washed with running water for about 10 hours. Wash the river sand and put it in flower pots of uniform size for later use. Then sow the washed seeds evenly in flower pots, sow 8 seeds in each pot, place them in the greenhouse, and water them once a day until they emerge. After the seedlings emerged, they wer...

Embodiment 3

[0128] Example 3: Acquisition of Arabidopsis SbbHLH85 Overexpression Lines and AtbHLH85 Mutant Lines

[0129] 1 Experimental materials

[0130] Select Columbia ecotype (Col-0) Arabidopsis and bHLH85 (RSL2 / AT4G33880) mutant Arabidopsis, T-DNA insertion mutants purchased from Tair website include rsl2-1 (SALK_048849), rsl2-2 (SALK_048857) , rsl2-3 (SALK_101872), rsl2-4 (SALK_143203).

[0131] 2 Experimental methods

[0132] 2.1 Obtaining Arabidopsis SbbHLH85 Overexpression Line

[0133] After the inflorescences of Arabidopsis thaliana were infiltrated with Agrobacterium pROKⅡ-SbbHLH85-GFP, the inflorescences of Arabidopsis thaliana were wrapped with black plastic wrap and cultured in the dark for about 24 hours, and then the black plastic wrap was uncovered for normal cultivation. After about 7 days, the second infection was carried out, the infection time was 10s, and the infection was continued for 3 times. The Arabidopsis thaliana to be infected matured and set seeds, and...

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Abstract

The invention provides the application of the anti-stress related protein bHLH85 in regulating the stress resistance of plants, and belongs to the field of biotechnology. The present invention reports for the first time a salt resistance negative regulation related protein bHLH85 and its coding gene. The present invention finds through research that the expression level of bHLH85 gene in sweet sorghum is significantly down-regulated under salt stress conditions, and it is introduced into Arabidopsis thaliana, and its functional identification shows that the overexpression of bHLH85 leads to the number and length of plant root hairs Increased, so that the salt-resistant ability of the plant decreased, which showed that it played a negative regulatory role in the salt-resistant ability of the plant, thus providing a basis for the cultivation of resistant plants.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the application of stress-resistance-related protein bHLH85 in regulating stress resistance of plants. Background technique [0002] The information disclosed in this background section is only intended to increase the understanding of the general background of the present invention, and is not necessarily taken as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to those skilled in the art. [0003] For a long time, soil salinization has been one of the main problems facing our country, which threatens many industries such as agriculture and animal husbandry. Salt stress has a significant impact on the growth and development of plants, and has an impact on physiological indicators such as plant growth rate, plant height, and biomass. Under the condition of salt stress, the initial manifestations are slow growth, ye...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/11C12N15/82A01H5/00A01H5/10A01H6/82A01H6/20
CPCC07K14/415C12N15/8218C12N15/8273C12N15/8271
Inventor 隋娜吴凤慧郑洪祥
Owner SHANDONG NORMAL UNIV
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